PROJECT SUMMARY Enzyme by Design, Inc. (EbD) is developing a safer version of L-asparaginase that can be used routinely in the treatment of adult ALL patients whose hypersensitivity to today's bacterial L-asparaginases deprives them from this efficacious therapy. EbD's L-asparaginase is novel due to its mammalian (guinea pig) origin and its lack of L-glutaminase co-activity. The need for better therapeutics for adult ALL is clear; the cure rate is <40%, and 80% of the deaths from ALL involve adults. In the US, the standard of care is the PEGylated E. coli enzyme (Oncaspar). Patients who develop complications with this 1st-line agent are moved to the L- asparaginase from E. chrysanthemi (Erwinaze). Issues that plague both drugs are their highly immunogenic nature (due to being of bacterial origin, though PEGylation decreases but not eliminates this challenge) and their L-glutaminase co-activity. The latter is source of many of the toxicities observed in patients, such as hepatotoxicity, pancreatitis, coagulation abnormalities and neurotoxicity. The manufacturers of both of these approved L-asparaginases are currently developing newer versions. However, those newer versions will still have the same toxicity issues since the underlying causes, their bacterial origin and L-glutaminase co-activity, remain in the products in development. In contrast, EbD's mammalian enzyme that is devoid of the L- glutaminase co-activity promises superiority by having reduced toxicity. In Aim 1 of the work supported by the STTR grant, EbD is finalizing the site-specific PEGylation strategy for GpA in order to achieve full masking of the enzyme by the PEG. Aim 2 will identify the PEGylated variants with the best pharmacokinetic (PK) properties, which will proceed to Aim 3 where it will be benchmarked against Oncaspar. Our data predicts that EbD's PEGylated GpA will exhibit significantly reduced toxicity compared to the current 1st-line treatment. Such a finding will provide the rationale for advancement to a STTR Phase II, where we will perform the pre-clinical studies required for an IND application. Our proposed I-CORPs team will include Dr. Arnon Lavie (founder & CEO), Mr. Stephen Morales (MBA, industry expert), and Dr. Hien Anh Nguyen (PI of STTR grant, founder & CSO). All three I-CORPs team members are committed to the time and effort requirements of the program. EbD has already conducted background research into the competitive landscape of the technology and identified the most commercially viable indication for the launch of the product. During the execution of the I- CORPS program, we plan to conduct an in-depth analysis with health care providers and payers to prepare to answer some of the key questions that angel/venture investors may have. The team expects to modify/refine the overall commercialization strategy based on the knowledge gained during the I-CORPS program.